A tracking technique for comparing individual home ranges of small mammals

The tracking technique consists of feeding marker bait to small mammals whilst in captivity, releasing them, and subsequently collecting and examining faeces deposited by them in containers in the study area. This technique allows a large number of individually marked animals to be tracked simultaneously over a short period of time, without restricting the movements of the animals in any way. This provides estimates of home range size that are considered to be more realistic than previous estimates, and allows the dynamic aspect of home range to be measured.     

A technique for quantitative estimation of small mammals traversing water obstacles

A new method for quantitative study of small mammals swimming across water obstacles was developed. A line of 25 traps was installed on either poles or boards (“rafts”) with anchors at a distance of 20–25 m from the bank and with 10-m distances between the items. The study was performed upstream along the Ilych River in August 2013. A total of 300 trap/day were accumulated. Twenty-four small mammal individuals of 8 species were captured. Their relative abundance was estimated as the number of individuals per 100 trap/day. It was found experimentally that floating poles neither repel nor attract animals. When an individual accidentally finds a floating pole, it climbs up and explores it for some time. The number of animals per total length of rafts per time unit can be suggested as an index of intensiveness of migration across a water obstacle. In the area studied, the number of small mammals of various species crossing the river was estimated at 26.7 individuals per 1 km/day. A length of 5 m for floating poles/boards and installation of two traps at the ends of an item is suggested to be used.     

A new version of the Ag?NOR technique. A combination with DAPI staining

Summary The Ag−NOR staining technique is widely used for visualizing nucleolar organizer regions (NORs) in various plant and animal tissues. We describe a simple and time-saving combination of Ag−NOR staining with DNA detection by fluorescence microscopy. This modification was tested on cultured cells and semi-thin sections of plastic-embedded tissues. Of the different fixatives and embedding media used in our studies, the best results (i.e., high selectivity of staining, and lack of or very low background precipitation) were obtained with fixation in methanol-acetone at −20°C for cultured cells, and fixation in 4% formaldehyde followed by embedding in Histocryl resin for tissue sections. The optimal time of Ag−NOR staining was determind experimentally for all materials tested. The specificity of the staining was checked at the electron microscopical level. Especially good results were obtained by mixing epifluorescence with standard bright-field illumination. In such a combination, Ag−NOR-positive nucleoli, or their fibrillar centres and dense fibrillar components, were clearly visible against a bright background of nuclear DNA.     

A new staining technique for microfilariae.

A rapid whole mount staining method is described to identify and differentiate microfilariae without elaborate processing. A single solution combining Hoyer's mounting medium and hematoxylin stain facilitates light microscopic examination of nuclei and sheaths of microfilariae. The new technique stains microfilariae adequately in three to seven minutes at 60--64 C making the method preferable to conventional methods that may take as long as 45 to 60 minutes. Lantern heat may be used to heat slides in rural areas with good results.     

A novel technique for immunohistoperoxidase staining of unfixed whole joints of small animals

Summary A method has been developed to cut unfixed and undecalcified sections of rat paws from animals with adjuvant arthritis and to stain them by a biotin-avidin immunoperoxidase technique. Good tissue integrity and morphology throughout the immunohistochemical procedure were retained if the sections were first mounted on transparent sellotape. The method is illustrated with two monoclonal antibodies (mAb) and is generally applicable with any mAb or polyclonal antibody and with joints from other small animals. For rats with adjuvant arthritis it was found important to block endogenous peroxidase before immunostaining. Complete inhibition of this enzyme without loss of antigenicity was best achieved after application of the mAb and biotinylated anti-IgG conjugate to the unfixed tissue sections.     

A modified mordant technique for staining plant chromosomes.

The ability to produce ferric acetate in house has provided a new mordant source for use in Newcomer's fluid (6:3:1:1:1 isopropanol, propionic acid, acetone, petroleum ether, 1,4-dioxane) for species with small chromosomes. This method improves on one first published in 1970.     

A hybrid technique for staining meiotic nuclei of Basidiomycetes.

A hybrid nuclear staining procedure used on the basidiomycete Schizophyllum commune produced squash preparations of sufficient density for effective brightfield photomicrography. Procedure: Tissure is fixed in Newcomer's solution, rehydrated in a graded ethanol series, rinsed, hydrolyzed in 1 N HC1 at 60 C, washed, mordanted in 4% iron alum, rinsed again, and stained with aceto-iron-hematoxylin prior to mounting and squashing.     

A new fluorescence technique for staining of mononuclear phagocytes.

A recently developed fluorochrome, 4',6-diamidino-2-phenylindole (DAPI), is used to stain mononuclear phagocytes of the mouse. After addition of heparin, these cells showed a bright yellow outer ring.     

Movement distances and density estimation of small mammals using the spool-and-line technique

Edge effect is an inherent problem when using trapping grids to estimate density of small mammals, resulting in a sampling area larger than the area of the grid. Distances between captures of individuals are used to estimateA(W), the effective sampling area of a trapping grid, but grid size sets a limit for the largest detectable distance. The spool-and-line technique is proposed here as a new method to estimateA(W). Movement distances based on the spool-and-line technique were compared to similar movement distances based on capture-recapture of three species of marsupials of the Atlantic Forest of Brazil. Distances based on the two methods were uncorrelated, and only ln-transformed distances based on the spool-and-line were normally distributed. The maximum distance moved (MaxD) estimated by the spool-and-line was chosen as the more accurate and practical distance to estimate edge effect. Estimates of the effective sampling area and densities for the common opossumDidelphis aurita (Wied-Neuwied, 1826), were compared using MaxD based on spool-and-line (MaxDs p o o l ) , capture-recapture (MaxD_cap), and also the distance between traps (DT). MaxD_spool reflected more accurately density variation between seasons. Movement distances of small mammals based on the spool-and-line technique permit more accurate estimates of density and its dynamics.     

A sensitive direct calorimeter for small mammals

A sensitive direct calorimeter for small animals is presented. Its principle is based on the measurement of the heat transfer from the animal chamber to a heat sink. The system gives repetitive measurements with a high efficiency and allows a detailed time-related measurement of the heat production by the whole animal. Its low response time can be advantageously used for the study of post-prandial heat generation and diet-induced thermogenesis. Data on the heat production by Wistar and lean and obese Zucker rats is also included.     

Microwave-assisted technology for the clearing and staining of arbuscular mycorrhizal fungi in roots

The use of microwave irradiation as a source of energy to clear and stain intra-radical arbuscular mycorrhizal fungi propagules has been tested on a variety of indigenous and cultivated herbaceous plants. The aim of the study was to evaluate the efficiency of microwave irradiation on root softening, fungi tissue staining, and preservation of DNA integrity for subsequent molecular analyses. The proposed methodology has been adapted from the standard procedures used to detect and quantify mycorrhizal root colonization levels. Using a domestic microwave oven, tissue clearing and staining required together between 30 s and 1.5 min of microwave treatment to be completed, depending the diameter size of the roots. The well-performing chemical stains tested were acid fuchsin, trypan blue, and aniline blue. The acid fuchsin clearing and staining processes, as performed, were also demonstrated to preserve DNA integrity for further molecular analyses. Irradiation by microwaves has been used with success in our laboratory within the frame of several studies. It offers considerable time saving over traditional method, reducing processing times from several hours to a few minutes while decreasing considerably the amount of chemicals and energy required to perform analyses.